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Allele specific LAMP- gold nanoparticle for characterization of single nucleotide polymorphisms

Due to their relevance as disease biomarkers and for diagnostics, screening of single nucleotide polymorphism (SNPs) requires simple and straightforward strategies capable to provide results in medium throughput settings. Suitable approaches relying on isothermal amplification techniques have been e...

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Autores principales: Carlos, Fábio Ferreira, Veigas, Bruno, Matias, Ana S., Doria, Gonçalo, Flores, Orfeu, Baptista, Pedro V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5671399/
https://www.ncbi.nlm.nih.gov/pubmed/29124021
http://dx.doi.org/10.1016/j.btre.2017.10.003
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author Carlos, Fábio Ferreira
Veigas, Bruno
Matias, Ana S.
Doria, Gonçalo
Flores, Orfeu
Baptista, Pedro V.
author_facet Carlos, Fábio Ferreira
Veigas, Bruno
Matias, Ana S.
Doria, Gonçalo
Flores, Orfeu
Baptista, Pedro V.
author_sort Carlos, Fábio Ferreira
collection PubMed
description Due to their relevance as disease biomarkers and for diagnostics, screening of single nucleotide polymorphism (SNPs) requires simple and straightforward strategies capable to provide results in medium throughput settings. Suitable approaches relying on isothermal amplification techniques have been evolving to substitute the cumbersome and highly specialized PCR amplification detection schemes. Nonetheless, identification of an individual’s genotype still requires sophisticated equipment and laborious methods. Here, we present a low-cost and reliable approach based on the allele specific loop-mediated isothermal amplification (AS-LAMP) coupled to ssDNA functionalized gold nanoparticle (Au-nanoprobe) colorimetric sequence discrimination. The Au-nanoprobe integration allows for the colorimetric detection of AS-LAMP amplification product that can be easily interpreted in less than 15 min. We targeted a clinical relevant SNP responsible for lactose intolerance (-13910C/T dbSNP rs#: 4988235) to demonstrate its proof of concept and full potential of this novel approach.
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spelling pubmed-56713992017-11-09 Allele specific LAMP- gold nanoparticle for characterization of single nucleotide polymorphisms Carlos, Fábio Ferreira Veigas, Bruno Matias, Ana S. Doria, Gonçalo Flores, Orfeu Baptista, Pedro V. Biotechnol Rep (Amst) Article Due to their relevance as disease biomarkers and for diagnostics, screening of single nucleotide polymorphism (SNPs) requires simple and straightforward strategies capable to provide results in medium throughput settings. Suitable approaches relying on isothermal amplification techniques have been evolving to substitute the cumbersome and highly specialized PCR amplification detection schemes. Nonetheless, identification of an individual’s genotype still requires sophisticated equipment and laborious methods. Here, we present a low-cost and reliable approach based on the allele specific loop-mediated isothermal amplification (AS-LAMP) coupled to ssDNA functionalized gold nanoparticle (Au-nanoprobe) colorimetric sequence discrimination. The Au-nanoprobe integration allows for the colorimetric detection of AS-LAMP amplification product that can be easily interpreted in less than 15 min. We targeted a clinical relevant SNP responsible for lactose intolerance (-13910C/T dbSNP rs#: 4988235) to demonstrate its proof of concept and full potential of this novel approach. Elsevier 2017-10-27 /pmc/articles/PMC5671399/ /pubmed/29124021 http://dx.doi.org/10.1016/j.btre.2017.10.003 Text en © 2017 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Carlos, Fábio Ferreira
Veigas, Bruno
Matias, Ana S.
Doria, Gonçalo
Flores, Orfeu
Baptista, Pedro V.
Allele specific LAMP- gold nanoparticle for characterization of single nucleotide polymorphisms
title Allele specific LAMP- gold nanoparticle for characterization of single nucleotide polymorphisms
title_full Allele specific LAMP- gold nanoparticle for characterization of single nucleotide polymorphisms
title_fullStr Allele specific LAMP- gold nanoparticle for characterization of single nucleotide polymorphisms
title_full_unstemmed Allele specific LAMP- gold nanoparticle for characterization of single nucleotide polymorphisms
title_short Allele specific LAMP- gold nanoparticle for characterization of single nucleotide polymorphisms
title_sort allele specific lamp- gold nanoparticle for characterization of single nucleotide polymorphisms
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5671399/
https://www.ncbi.nlm.nih.gov/pubmed/29124021
http://dx.doi.org/10.1016/j.btre.2017.10.003
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